UHPLC-MS/MS for plasma lamotrigine analysis and comparison with a homogenous enzyme immunoassay.

Aims: To develop and validate a UHPLC-MS/MS method for lamotrigine (LTG) analysis in human plasma and evaluate its agreement with a homogenous enzyme immunoassay (HEIA). Materials & methods: The UHPLC-MS/MS method was developed and validated according to the USFDA/EMA guidelines. A Bland-Altman plot was used to evaluate the agreement between UHPLC-MS/MS and HEIA. Results: Samples were pretreated with one-step protein precipitation and separated in 2.6 min. The intra- and inter-day bias and imprecisions were -15.8 to 15.0% and less than 11.17%, respectively. The recovery and matrix factor were 98.30 to 111.97%. The mean overestimation of UHPLC-MS/MS compared with HEIA was 21.57%. Conclusion: A rapid, sensitive and robust UHPLC-MS/MS method for plasma LTG analysis was developed and validated and was a 21.57% overestimation compared with HEIA.

In Vitro Effect of Low-Level Lasers on Total Bilirubin Concentration in Human Blood Plasma Using 375 and 650 nm Lasers.

Background: The current research extends previous laboratory investigations by investigating the effects of low-level laser irradiation (LLLI) on human blood plasma. Total bilirubin is of special importance because of its potential biostimulatory and modulatory actions. Objective: This study aims to analyze changes in total bilirubin content as a consequence of LLLI on human blood plasma. This study aims to determine how changes in exposure duration and laser wavelength affect these adjustments. Methodology: Plasma was isolated from a healthy adult donor's whole blood using the anticoagulant ethylenediaminetetraacetic acid (EDTA). Plasma samples were exposed to LLLI at 375 and 650 nm for 5, 10, 15, 20, and 25 min. Total bilirubin concentrations were measured both before and after irradiation using spectrophotometric analysis. The difference between 375 and 630 nm lasers was also investigated. Results: Five, 10, 15, 20, and 25 min of exposure to LLLI at 375 and 650 nm wavelengths resulted in statistically significant differences in total bilirubin content (p ˂ 0.05, p ˂ 0.001, p ˂ 0.0001). There was no statistically significant difference in total bilirubin concentration between the 375 and 630 nm lasers. Conclusions: Human blood plasma total bilirubin levels were considerably lower following LLLI at 375 and 630 nm than controls. Multiple exposures provide the same results. These findings demonstrate the role of biostimulation by laser irradiation in blood plasma applications and suggest that low-level laser treatment may control total bilirubin levels, particularly at 375 and 630 nm.

Developing magnetic functionalized dendritic fibrous mesoporous silica as advanced adsorbent for quaternary ammonium alkaloids.

A novel π-conjugated polymer-modified magnetic dendritic fibrous mesoporous silica adsorbent (MB@KCC-1@π-CP) is reported for the accurate determination of quaternary ammonium alkaloids (QAAs) in complex body fluid matrices. It is demonstrated that the magnetic dendritic fibrous mesoporous silica (MB@KCC-1) is an excellent carrier combining magnetism, high specific surface area, unique hierarchical pore structure, and fast mass transfer rate. The π-conjugated polymer (π-CP) can efficiently retain QAAs (berberine, coptisine, palmatine, jatrorrhizine) by multiple interactions. In addition, the adsorption kinetics and adsorption mechanism were also studied and discussed. Under optimized extraction conditions, MB@KCC-1@π-CP-based magnetic solid-phase extraction (MSPE) and high-performance liquid chromatography (HPLC) method affords a wide linear range (0.5-20000 ng mL-1), low limits of detection (0.2-2 ng mL-1), and satisfactory relative standard deviations (RSD) of inter-day (< 2.4%) and intra-day (< 3.1%) for QAAs. Trace QAAs in complex human blood plasma samples were successfully detected by the established method.

Age prediction from human blood plasma using proteomic and small RNA data: a comparative analysis.

Aging clocks, built from comprehensive molecular data, have emerged as promising tools in medicine, forensics, and ecological research. However, few studies have compared the suitability of different molecular data types to predict age in the same cohort and whether combining them would improve predictions. Here, we explored this at the level of proteins and small RNAs in 103 human blood plasma samples. First, we used a two-step mass spectrometry approach measuring 612 proteins to select and quantify 21 proteins that changed in abundance with age. Notably, proteins increasing with age were enriched for components of the complement system. Next, we used small RNA sequencing to select and quantify a set of 315 small RNAs that changed in abundance with age. Most of these were microRNAs (miRNAs), downregulated with age, and predicted to target genes related to growth, cancer, and senescence. Finally, we used the collected data to build age-predictive models. Among the different types of molecules, proteins yielded the most accurate model (R² = 0.59 ± 0.02), followed by miRNAs as the best-performing class of small RNAs (R² = 0.54 ± 0.02). Interestingly, the use of protein and miRNA data together improved predictions (R2 = 0.70 ± 0.01). Future work using larger sample sizes and a validation dataset will be necessary to confirm these results. Nevertheless, our study suggests that combining proteomic and miRNA data yields superior age predictions, possibly by capturing a broader range of age-related physiological changes. It will be interesting to determine if combining different molecular data types works as a general strategy to improve future aging clocks.

Affinity-Purification Combined with Crosslinking Mass Spectrometry for Identification and Structural Modeling of Host-Pathogen Protein-Protein Complexes.

Host-pathogen protein-protein interactions are highly complex and dynamic and mediate key steps in pathogen adhesion to host, host invasion, and colonization as well as immune evasion. In bacteria, these interactions most often involve specialized virulence factors or effector proteins that specifically target central host proteins. Here, I present a mass spectrometry-based proteomics approach starting with the identification of host-pathogen interactions by affinity-purification followed by mapping the specific host-pathogen protein-protein interaction interfaces by crosslinking mass spectrometry and structural modeling of the complexes.

Zwitterionic Polysulfobetaine Coating and Antiplatelet Liposomes Reduce Fouling in Artificial Lung Circuits.

The artificial lung has provided life-saving support for pulmonary disease patients and recently afforded patients with severe cases of COVID-19 better prognostic outcomes. While it addresses a critical medical need, reducing the risk of clotting inside the device remains challenging. Herein, a two-step surface coating process of the lung circuit using Zwitterionic polysulfobetaine methacrylate is evaluated for its nonspecific protein antifouling activity. It is hypothesized that similarly applied coatings on materials integrated (IT) or nonintegrated (NIT) into the circuit will yield similar antifouling activity. The effects of human plasma preconditioned with nitric oxide-loaded liposome on platelet (plt) fouling are also evaluated. Fibrinogen antifouling activities in coated fibers are similar in the IT and NIT groups. It however decreases in coated polycarbonate (PC) in the IT group. Also, plt antifouling activity in coated fibers is similar in the IT and NIT groups and is lower in coated PC and Tygon in the IT group compared to the NIT group. Coating process optimization in the IT lung circuit may help address difference in the coating appearance of outer and inner fiber bundle fibers, and the NO-liposome significantly reduces (86%) plt fouling on fibers indicating its potential use for blood anticoagulation.

Enhancement of microsphere specificity to purify human serum albumin from blood plasma.

Efforts to rapidly and easily supply purified human serum albumin (HSA) in remote areas during a pandemic are challenging. Here, we developed an HSA-imprinted microsphere (HSAIM) as a matrix to purify HSA from blood plasma on a small scale. HSAIMs were generated by encapsulating silica-3-(2-imidazoline-1-yl) propyltriethoxysilane-Cu2+-HSA (SiO2-IMEO-Cu2+-HSA) into agarose gel, and the stereospecific template for HSA was obtained by eluting the agarose gel. The physicochemical properties and performance of HSAIM were evaluated, and HSAIMs were applied to purify HSA from human blood plasma. Spherical HSAIMs had an average diameter of 51.2 ± 6.1 µm. HSAIMs had a maximum adsorption of 8.77 × 10-2 µmol HSA g-1 with an imprinting factor of 2.83, and the selectivity factors of BSA, thrombin and IgG were 0.96, 0.59 and 0.26, respectively. HSAIMs had a dynamic binding capacity (DBC10%) of 5.94 × 10-2 µmol HSA g-1 and could be reused up to 10 cycles with an ultimate recovery of 55.92%. HSA adsorption kinetics of HSAIM fitted to a pseudo-second-order mechanism, and HSA binding characteristics fit with a Sips isotherm model. For practical purposes, an initial blood plasma sample containing 24.9 ± 2.5 mg protein was pretreated with ethanol yielding 14.5 ± 4.6 mg protein, and further purification with HSAIM yielded 3.6 ± 1.1 mg protein. Starting with a blood plasma sample containing 149 type proteins, a single protein identified as HSA was obtained after final purification step with the HSAIM column, indicating that HSAIMs stereospecifically bound HSA. Hence, HSAIM was promising for blood plasma purification on a small scale.

Core-shell bioprinting of vascularizedin vitroliver sinusoid models.

In vitroliver models allow the investigation of the cell behavior in disease conditions or in response to changes in the microenvironment. A major challenge in liver tissue engineering is to mimic the tissue-level complexity: besides the selection of suitable biomaterial(s) replacing the extracellular matrix (ECM) and cell sources, the three-dimensional (3D) microarchitecture defined by the fabrication method is a critical factor to achieve functional constructs. In this study, coaxial extrusion-based 3D bioprinting has been applied to develop a liver sinusoid-like model that consists of a core compartment containing pre-vascular structures and a shell compartment containing hepatocytes. The shell ink was composed of alginate and methylcellulose (algMC), dissolved in human fresh frozen plasma. The algMC blend conferred high printing fidelity and stability to the core-shell constructs and the plasma as biologically active component enhanced viability and supported cluster formation and biomarker expression of HepG2 embedded in the shell. For the core, a natural ECM-like ink based on angiogenesis-supporting collagen-fibrin (CF) matrices was developed; the addition of gelatin (G) enabled 3D printing in combination with the plasma-algMC shell ink. Human endothelial cells, laden in the CFG core ink together with human fibroblasts as supportive cells, formed a pre-vascular network in the core in the absence and presence of HepG2 in the shell. The cellular interactions occurring in the triple culture model enhanced the albumin secretion. In conclusion, core-shell bioprinting was shown to be a valuable tool to study cell-cell-interactions and to develop complex tissue-like models.

Towards Label-free detection of viral disease agents through their cell surface proteins: Rapid screening SARS-CoV-2 in biological specimens.

Current methods for the screening of viral infections in clinical settings, such as reverse transcription polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay (ELISA), are expensive, time-consuming, require trained personnel and sophisticated instruments. Therefore, novel sensors that can save time and cost are required specially in remote areas and developing countries that may lack the advanced scientific infrastructure for this task. In this work, we present a sensitive, and highly specific biosensing approach for the detection of harmful viruses that have cysteine residues within the structure of their cell surface proteins. We utilized new method for the rapid screening of SARS-CoV-2 virus in biological fluids through its S1 protein by surface enhanced Raman spectroscopy (SERS). The protein is captured from aqueous solutions and biological specimens using a target-specific extractor substrate. The structure of the purified protein is then modified to convert it into a bio-thiol by breaking the disulfide bonds and freeing up the sulfhydryl (SH) groups of the cysteine residues. The formed biothiol chemisorbs favourably onto a highly sensitive plasmonic sensor and probed by a handheld Raman device in few seconds. The new method was used to screen the S1 protein in aqueous medium, spiked human blood plasma, mucus, and saliva samples down to 150 fg/L. The label-free SERS biosensing method has strong potential for the fingerprint identification many viruses (e.g. the human immunodeficiency virus, the human polyomavirus, the human papilloma virus, the adeno associated viruses, the enteroviruses) through the cysteine residues of their capsid proteins. The new method can be applied at points of care (POC) in remote areas and developing countries lacking sophisticated scientific infrastructure.

Protocol for Measuring Concentrations of Extracellular Vesicles in Human Blood Plasma with Flow Cytometry.

Extracellular vesicles (EVs) are lipid membrane enclosed particles that are released from cells into body fluids, such as blood. EVs offer potential new biomarkers of diseases, because the cellular origin, composition, concentration, and function of EVs change in health and disease. The concentration of EVs from specific cell types in blood can be determined with flow cytometry. A flow cytometer measures fluorescence and light scattering signals from single EVs, but only if these signals are sufficiently bright to be detected. Measured concentrations of EVs are therefore only reproducible and comparable if the detection ranges are known and reported in standard units, such as molecules of equivalent soluble fluorophore (MESF) for fluorescence signals and the diameter in nm for scatter signals. The goal of this protocol is to discuss all steps needed to derive the concentration of cell-type specific EVs within a known diameter range and fluorescence range. More specifically, this protocol describes how to determine the concentration of CD61+ (Integrin beta-3, platelet marker), CD235a+ (Glycophorin A, erythrocyte marker), and CD45+ (leukocyte common antigen) EVs in human blood plasma with an Apogee A60-Micro flow cytometer using scatter-based triggering. The principles behind this protocol could lay a firm basis for the design of a protocol suitable for other flow cytometers and body fluids.

Highly selective and sensitive detection of glutathione over cysteine and homocysteine with a turn-on fluorescent biosensor based on cysteamine-stabilized CdTe quantum dots.

In this work, cysteamine (CA) stabilized CdTe quantum dots (QDs) (CA-CdTe QDs) and sodium citrate stabilized gold nanoparticles (AuNPs) were prepared. Because of the strong electrostatic interaction and spectral overlap of emission spectrum of CA-CdTe QDs and absorption spectrum of AuNPs, a highly effective fluorescence resonance energy transfer (FRET) system was formed and the fluorescence of CA-CdTe QDs was strongly quenched. The synthesized CA-CdTe and AuNPs were self-assembled to large clusters due to the electrostatic attraction and the fluorescence of CA-CdTe was sharply quenched as a result of FRET. Under the optimum pH of 5.5, the positive GSH could assemble with negative AuNPs through electrostatic interaction and destroy the FRET system of CA-CdTe and AuNPs, due to the red shift of absorption wavelength of AuNPs caused by aggregation. The fluorescence of CA-CdTe recovered, and the recovered fluorescence efficiency shows a linear function against the GSH concentrations from 6.7 nM to 0.40 µM, with a detecting limit of 3.3 nM. The quenched emission of CA-CdTe could be recovered attributed to the aggregation of AuNPs by GSH. Under optimal conditions, the sensing system was successfully applied in the detection of GSH in real human blood plasma samples with a recovery of 99.5-102.3%, showing a promising future for the highly sensitive and selective GSH detection in the human blood plasma samples.

Neuroprotection Mediated by Human Blood Plasma in Mouse Hippocampal Slice Cultures and in Oxidatively Stressed Human Neurons.

Neuroprotection from oxidative stress is critical during neuronal development and maintenance but also plays a major role in the pathogenesis and potential treatment of various neurological disorders and neurodegenerative diseases. Emerging evidence in the murine system suggests neuroprotective effects of blood plasma on the aged or diseased brain. However, little is known about plasma-mediated effects on human neurons. In the present study, we demonstrate the neuroprotective effect mediated by human plasma and the most abundant plasma-protein human serum albumin against oxidative stress in glutamatergic neurons differentiated from human neural crest-derived inferior turbinate stem cells. We observed a strong neuroprotective effect of human plasma and human serum albumin against oxidative stress-induced neuronal death on the single cell level, similar to the one mediated by tumor necrosis factor alpha. Moreover, we detected neuroprotection of plasma and human serum albumin against kainic acid-induced excitatory stress in ex vivo cultured mouse hippocampal tissue slices. The present study provides deeper insights into plasma-mediated neuroprotection ultimately resulting in the development of novel therapies for a variety of neurological and, in particular, neurodegenerative diseases.

Gold nanoparticle enhanced multiplexed biosensing on a fiber optic surface plasmon resonance probe.

We present an innovative multiplexing concept on a fiber optic surface plasmon resonance (FO-SPR) platform and demonstrate for the first time the simultaneous detection of two targets using the same FO sensor probe. Co(III)-NTA chemistry was used for oriented and stable co-immobilization of two different His6-tagged bioreceptors. T2C2 and MDTCS (i.e. fragments of the ADAMTS13 metalloprotease linked to the thrombotic thrombocytopenic purpura disorder) served as model system bioreceptors together with their respective targets (4B9 and II-1 antibodies). Gold nanoparticles were used here in an original way for discriminating the two targets in the same sample, in addition to their traditional signal amplification-role. After verifying the specificity of the selected model system, we studied the bioreceptor surface density and immobilization order. Innovative approach to lower the bioreceptor concentration below surface saturation resulted in an optimal detection of both targets, whereas the order of immobilization of the two bioreceptors did not give any significant difference. By sequentially immobilizing the T2C2 and MDTC bioreceptors, we established calibration curves in buffer and 100-fold diluted human blood plasma. This resulted in calculated limits of detection of 3.38 and 2.31 ng/mL in diluted plasma for 4B9 and II-1, respectively, indicating almost the same sensitivity as in buffer. Importantly, we also proved the applicability of the established calibration curves for quantifying the targets at random and more realistic ratios, directed by the design of experiments. This multiplexing study further expands the repertoire of applications on the FO-SPR biosensing platform, which together with its intrinsic features opens up great opportunities for diagnostics and life sciences.

miREV: An Online Database and Tool to Uncover Potential Reference RNAs and Biomarkers in Small-RNA Sequencing Data Sets from Extracellular Vesicles Enriched Samples.

Extracellular vesicles (EVs) are nano-sized, membrane-enclosed vesicles released by cells for intercellular communication. EVs are involved in pathological processes and miRNAs in EVs have gained interest as easily accessible biomolecules in liquid biopsies for diagnostic purposes. To validate potential miRNA biomarker, transcriptome analyses must be carried out to detect suitable reference miRNAs. miREV is a database with over 400 miRNA sequencing data sets and helps the researcher to find suitable reference miRNAs for their individual experimental setup. The researcher can put together a specific sample set in miREV, which is similar to his own experimental concept in order to find the most suitable references. This allows to run validation experiments without having to carry out a complex and costly transcriptome analysis priorly. Additional read count tables of each generated sample set are downloadable for further analysis. miREV is freely available at https://www.physio.wzw.tum.de/mirev/.

2D metal azolate framework as nanozyme for amperometric detection of glucose at physiological pH and alkaline medium.

The synthesis of Co-based two-dimensional (2D) metal azolate framework nanosheets (MAF-5-CoII NS) is described using a simple hydrothermal method. The product was isostructural to MAF-5 (Zn). The as-prepared MAF-5-CoII NS exhibited high surface area (1155 m2/g), purity, and crystallinity. The MAF-5-CoII NS-modified screen-printed electrode (MAF-5-CoII NS/SPE) was used for nonenzymatic detection of glucose in diluted human blood plasma (BP) samples with phosphate buffer saline (PBS, pH 7.4) and NaOH (0.1 M, pH 13.0) solutions. The MAF-5-CoII NS nanozyme displayed good redox activity in both neutral and alkaline media with the formation of CoII/CoIII redox pair, which induced the catalytic oxidation of glucose. Under the optimized detection potential, the sensor presented a chronoamperometric current response for the oxidation of glucose with two wide concentration ranges in PBS-diluted (62.80 to 180 µM and 305 to 8055 µM) and NaOH-diluted (58.90 to 117.6 µM and 180 to 10,055 µM) BP samples, which were within the limit of blood glucose levels of diabetic patients before (4.4-7.2 mM) and after (10 mM) meals (recommended by the American Diabetes Association). The sensor has a limit of detection of ca. 0.25 and 0.05 µM, respectively, and maximum sensitivity of ca. 36.55 and 1361.65 mA/cm2/mM, respectively, in PBS- and NaOH-diluted BP samples. The sensor also displayed excellent stability in the neutral and alkaline media due to the existence of hydrophobic linkers (2-ethyl imidazole) in the MAF-5-CoII NS, good repeatability and reproducibility, and interference-free signals. Thus, MAF-5-CoII NS is a promising nanozyme for the development of the disposable type of sensor for glucose detection in human body fluids. Graphical abstract.

Identification of 2-(diethylamino)ethylthiol dipeptide (Cys-Pro) adduct as biomarker of nerve agents VR and CVX in human plasma using liquid chromatography-high-resolution tandem mass spectrometry.

Organophosphorus nerve agents pose a significant threat to human health. The most toxic compounds in this class include V-type poisonous substances such as VX, CVX, and VR. Although all stockpiles of this type of substance are subject to destruction under the Chemical Weapons Convention (CWC), there is still a risk that they could be used for criminal and terrorist purposes. The latter determines the relevance of studies aimed at identification of biomarkers that may indicate the exposure of these group substances to the organism. A liquid chromatography mass spectrometry/high-resolution mass spectrometry (LC-MS/HR MS) method for determination of trace amounts of nerve agents such as VR and CVX in human plasma was proposed. The method is based on enzymatic plasma hydrolysis with the use of pronase to form a stable adduct of 2-(diethylamino)ethylthiol with dipeptide cysteine-proline (DEAET-CP) with its subsequent determination by LC-MS/HR MS. Synthesis of DEAET-CP as reference compound was conducted using non-toxic precursors. Sample preparation of human blood plasma samples exposed to VR was carried out with the use of solid-phase extraction (SPE). Liquid chromatography (LC) separation on the reversed-phase column and mass spectrometric detection (selection of optimal transitions and detection modes) were performed. The achieved limit of detection (LOD) of VR (in the form of DEAET-CP) in human blood plasma was 0.05 ng mL-1. The proposed approach was developed using plasma samples exposed to VR and CVX obtained in the frame of the Fifth Official Biomedical Test of the Organization for the Prohibition of Chemical Weapons (OPCW) and showed good specificity of detection.

Doxorubicin delivery systems based on doped CaCO3 cores and polyanion drug conjugates.

In order to prolong the release and reduce the toxicity of anticancer drug - doxorubicin (DOX), delivery systems (DS) using different polyanions have been developed. Structural (size, morphological stability) and functional (encapsulation efficiency, DOX release) characteristics of three types of DS are compared: CaCO3 porous vaterites doped with polyanions by co-precipitation and coating techniques, and DOX-polyanion conjugates. Using scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS), it was shown that the doping enhances the morphological stability of CaCO3-based DS during the DOC loading. Doping of CaCO3 cores by co-precipitation reduces its sizes (up to 1 µm) and DOX encapsulation efficiency. Polyanion-coated CaCO3 cores and polyanion drug conjugates show about 98 w/w% DOX encapsulation. For the first time, it was shown that the release of DOX from developed DS into human blood plasma is more intense (from 1.3 to 3.0 times for different DS) than into model tumour environment.

Confirmation and Identification of Biomarkers Implicating Environmental Triggers in the Pathogenesis of Type 1 Diabetes.

Multiple environmental triggers have been proposed to explain the increased incidence of type 1 diabetes (T1D). These include viral infections, microbiome disturbances, metabolic disorders, and vitamin D deficiency. Here, we used ELISA to examine blood plasma from juvenile T1D subjects and age-matched controls for the abundance of several circulating factors relevant to these hypotheses. We screened plasma for sCD14, mannose binding lectin (MBL), lipopolysaccharide binding protein (LBP), c-reactive protein (CRP), fatty acid binding protein 2 (FABP2), human growth hormone, leptin, total adiponectin, high molecular weight (HMW) adiponectin, total IgG, total IgA, total IgM, endotoxin core antibodies (EndoCAbs), 25(OH) vitamin D, vitamin D binding protein, IL-7, IL-10, IFN-γ, TNF-α, IL-17A, IL-18, and IL-18BPa. Subjects also were tested for prevalence of antibodies targeting adenovirus, parainfluenza 1/2/3, Coxsackievirus, cytomegalovirus, Epstein-Barr virus viral capsid antigen (EBV VCA), herpes simplex virus 1, and Saccharomyces cerevisiae. Finally, all subjects were screened for presence and abundance of autoantibodies targeting islet cell cytoplasmic proteins (ICA), glutamate decarboxylase 2 (GAD65), zinc transporter 8 (ZNT8), insulinoma antigen 2 (IA-2), tissue transglutaminase, and thyroid peroxidase, while ß cell function was gauged by measuring c-peptide levels. We observed few differences between control and T1D subjects. Of these, we found elevated sCD14, IL-18BPa, and FABP2, and reduced total IgM. Female T1D subjects were notably elevated in CRP levels compared to control, while males were similar. T1D subjects also had significantly lower prevalence of EBV VCA antibodies compared to control. Lastly, we observed that c-peptide levels were significantly correlated with leptin levels among controls, but this relationship was not significant among T1D subjects. Alternatively, adiponectin levels were significantly correlated with c-peptide levels among T1D subjects, while controls showed no relationship between these two factors. Among T1D subjects, the highest c-peptide levels were associated with the lowest adiponectin levels, an indication of insulin resistance. In total, from our examination we found limited data that strongly support any of the hypotheses investigated. Rather, we observed an indication of unexplained monocyte/macrophage activation in T1D subjects judging from elevated levels of sCD14 and IL-18BPa. These observations were partnered with unique associations between adipokines and c-peptide levels among T1D subjects.

[The results of evaluating the degree of influence of human blood plasma on the strength of adhesion of composite material to dentin (one-component adhesive system)]. / Rezul'taty otsenki stepeni vliyaniya plazmy krovi cheloveka na prochnost' adgezionnogo soedineniya kompozitnogo materiala i dentina zuba (odnokomponentnaya adgezivnaya sistema).

OBJECTIVE: To study the dynamics of changes in the adhesion between the composite and dentin of the tooth when using different volumes of a single-component adhesive system in case of contamination of it with human blood plasma. MATERIAL AND METHODS: To achieve this goal, the following were used: extracted human teeth, XP Bond one-component adhesive systems (DENTSPLY, Germany), Esthet X HD micrometric restoration material (DENTSPLY, Germany), and centrifuged blood plasma. The strength of the adhesive bond between the composite material and the hard tissues of the tooth was studied using a shear test machine Zwick Roell Z 010 («Zwick¼, Germany). RESULTS: The use of a single-component adhesive system in an amount of 17.7 mg (1 drop from a dispenser) for treating open dentin makes it more resistant to contamination compared to using the same adhesive, but in an amount of 6.6 mg (the amount of adhesive that adsorbs a medium-sized dental take). A decrease in the adhesion force between the composite material and tooth hard tissues from 1.5 to 17.7% occurs when a single-component adhesive system weighing 17.7 mg of blood plasma simulating a dentinal fluid weighing from 0.2 to 2.0 mg enters. The ingestion of the same amount of blood plasma in a single-component adhesive system weighing 6.6 mg leads to a decrease in its adhesion from 4.3 to 43%.

Evaluation of lipid quantification accuracy using HILIC and RPLC MS on the example of NIST® SRM® 1950 metabolites in human plasma.

Lipidomics analysis for large-scale studies aiming at the identification and quantification of natural lipidomes is often performed using LC-MS-based data acquisition. However, the choice of suitable LC-MS method for accurate lipid quantification remains a matter of debate. Here, we performed the systematic comparison between two HRAM-MS-based quantification workflows based on HILIC and RPLC MS by quantifying 191 lipids from five lipid classes in human blood plasma using deuterated standards in the "one ISTD-per-lipid class" approach. Lipid quantification was performed considering all necessary isotopic corrections, and obtained correction factors are illustrated. Concentrations of lipids in NIST® SRM® 1950 human blood plasma determined by the two methods were comparable for most of the studied lipid species except for highly unsaturated phosphatidylcholines (PC). A comparison of lipid concentrations to consensus values determined in a previously published multi-laboratory study illustrated possible "overestimation" of concentrations for these highly unsaturated lipids by HILIC MS. We evaluated the influence of lipid loading amounts as well as the difference between quantified lipid and internal standard concentrations on the HILIC MS quantification results. We conclude that both HILIC and RPLC HRAM-MS workflows can be equally used for accurate lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), phosphatidylcholine (PC), phosphatidylethanolamine (PE), and sphingomyelin (SM) lipid quantification, despite significant differences in the concentration of highly unsaturated PC lipids which need to be addressed by establishing response factors to account for the differences in degree of lipid unsaturation. Graphical.